Mammalia

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    Protocols in Current Issue
    Analysis of Generalized Fibrosis in Mouse Tissue Sections with Masson’s Trichrome Staining
    Authors:  Diantha van de Vlekkert, Eda Machado and Alessandra d’Azzo, date: 05/20/2020, view: 698, Q&A: 0
    [Abstract] Expansion of fibrous connective tissue and abnormal deposition of extracellular matrix (ECM) are at the basis of many fibrotic diseases. Fibrosis can occur in response to both physiological and pathological cues, including wound healing, tissue remodeling/repair and inflammation. Chronic fibrosis can lead to severe tissue damage, organ failure and ...
    A Modified Semisolid Clonal Culture for Identification of B-1 and B-2 Progenitor Colony Forming Ability of Mouse Embryonic Hemogenic Endothelial Cells
    Authors:  Michihiro Kobayashi and Momoko Yoshimoto, date: 05/05/2020, view: 675, Q&A: 0
    [Abstract] The search for the origin of the first hematopoietic stem cells (HSCs) in the mouse embryo has been a hot topic in the field of developmental hematopoiesis. Detecting lymphoid potential is one of the supportive evidence to show the definitive hematopoietic activity of HSCs. However, the first B-lymphoid potential in the mouse embryos are reported ...
    Isolation, Purification and Characterization of Exosomes from Fibroblast Cultures of Skeletal Muscle
    Authors:  Diantha van de Vlekkert, Xiaohui Qiu, Ida Annunziata and Alessandra d'Azzo, date: 04/05/2020, view: 635, Q&A: 0
    [Abstract] Exosomes are dynamic nanovesicles secreted by virtually all cells and are present in all biological fluids. Given their highly heterogeneous content exosomes have been implicated in many physiological and pathological processes that they exert by influencing cell-cell and cell-ECM communication. In recent years an increasing number of methods have ...
    Lipid Mixing Assay for Murine Myoblast Fusion and Other Slow Cell-cell Fusion Processes
    [Abstract] Lipid mixing (redistribution of lipid probes between fusing membranes) has been widely used to study early stages of relatively fast viral and intracellular fusion processes that take seconds to minutes. Lipid mixing assays are especially important for identification of hemifusion intermediates operationally defined as lipid mixing without content ...
    RNA Sequencing of Single Myofibers from Mus musculus
    Authors:  Darren M. Blackburn, Felicia Lazure and Vahab D. Soleimani, date: 02/20/2020, view: 1683, Q&A: 0
    [Abstract] Whole transcriptome analysis is a key method in biology that allows researchers to determine the effect a condition has on gene regulation. One difficulty in RNA sequencing of muscle is that traditional methods are performed on the whole muscle, but this captures non-myogenic cells that are part of the muscle. In order to analyze only the ...
    Immunohistochemical Identification of Muscle Fiber Types in Mice Tibialis Anterior Sections
    Authors:  Vanitha V. Rao and Abhishek Mohanty, date: 10/20/2019, view: 1368, Q&A: 0
    [Abstract] Mammalian skeletal muscle is a metabolically active tissue that is made up of different types of muscle fibers. These myofibers are made up of important contractile proteins that provide force during contraction of the muscle like actin and myosin. Murine myofibers have been classified into 4 types: Type I, Type IIa, Type IIb and Type IIX. Each ...
    Isolation and Transcriptomic Profiling of Single Myofibers from Mice
    [Abstract] Skeletal muscle is composed of different cells and myofiber types, with distinct metabolic and structural features. Generally, transcriptomic analysis of skeletal muscle is performed using whole muscle, resulting in average information as all cells composing the organ contribute to the expression value detected for each gene with the loss of ...
    Isolation and Culture of Single Myofiber and Immunostaining of Satellite Cells from Adult C57BL/6J Mice
    Authors:  Shujie Chen, Hongrong Ding, Xiangping Yao and Liwei Xie, date: 07/20/2019, view: 2567, Q&A: 0
    [Abstract] Myofiber isolation followed with ex vivo culture could recapitulate and visualize satellite cells (SCs) activation, proliferation, and differentiation. This approach could be taken to understand the physiology of satellite cells and the molecular mechanism of regulatory factors, in terms of the involvement of intrinsic factors over SCs ...
    Cell Microencapsulation and Cryopreservation with Low Molecular Weight Hyaluronan and Dimethyl Sulfoxide
    Authors:  H. Gurruchaga, L. Saenz del Burgo, G. Orive, R. M. Hernandez, J. Ciriza and J. L. Pedraz, date: 02/20/2019, view: 2255, Q&A: 0
    [Abstract] Cryopreservation is commonly used for the storage of cells, tissues, organs or 3D cell-based products using ultra-low temperatures, which involves the immersion in liquid nitrogen for their long-term preservation. The cryopreservation of several microencapsulated cells is usually performed by the slow freezing with the dimethyl sulfoxide (DMSO) as ...
    Insulin Tolerance Test under Anaesthesia to Measure Tissue-specific Insulin-stimulated Glucose Disposal
    [Abstract] Insulin resistance is a pathophysiological state defined by impaired responses to insulin and is a risk factor for several metabolic diseases, most notably type 2 diabetes. Insulin resistance occurs in insulin target tissues including liver, adipose and skeletal muscle. Methods such as insulin tolerance tests and hyperinsulinaemic-euglycaemic ...



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